Nie Fang, Wang Xiao-Feng, Zhao Shi-Yi, Bu Lan, Liu Xue-Hui
Department of Medical Ultrasonics, Lanzhou University Second Hospital , Lanzhou, Gansu , China and.
J Drug Target. 2015 May;23(4):380-6. doi: 10.3109/1061186X.2014.1002500. Epub 2015 Feb 12.
The objective of this study was to assess the change of cytoskeleton and cell cycle in LoVo (human colorectal cancer) cell via gene silencing of Rac1 with RNA interference mediated by microbubble (SonoVue) and ultrasound (US).
The compound of plasmid Rac1-shRNA, LoVo cells, and SonoVue was exposed to US (1 MHz, 2 W/cm(2), 5 min). The expression of Rac1 mRNA and Rac1 protein was detected by RT-PCR and Western blot. Cytoskeleton was taken by confocal microscope in a random fashion. Cell invasion was assayed using modified Boyden chambers, and cell cycle and apoptosis were analyzed by flow cytometry in LoVo cells.
Rac1 gene is overexpressed in human colorectal cancer cells, gene silencing of Rac1 with RNA interference mediated by microbubble and US strongly inhibited lamellipodia formation, cell invasion, and delayed cell cycle, as well as enhanced cell apoptosis of LoVo cells in vitro.
Silencing Rac1 gene mediated by microbubble and US may become a new treatment option for the inhibition of the invasion and metastasis of colorectal cancer cells.
本研究旨在评估通过微泡（声诺维）和超声介导的RNA干扰使Rac1基因沉默后，洛沃（人结肠癌细胞）细胞中细胞骨架和细胞周期的变化。
将质粒Rac1-shRNA、洛沃细胞和声诺维的复合物暴露于超声（1兆赫，2瓦/平方厘米，5分钟）。通过RT-PCR和蛋白质印迹法检测Rac1 mRNA和Rac1蛋白的表达。用共聚焦显微镜随机观察细胞骨架。使用改良的博伊登小室检测细胞侵袭，并通过流式细胞术分析洛沃细胞的细胞周期和凋亡情况。
Rac1基因在人结肠癌细胞中过度表达，微泡和超声介导的RNA干扰使Rac1基因沉默可强烈抑制丝状伪足形成、细胞侵袭并延迟细胞周期，同时增强体外洛沃细胞的凋亡。
微泡和超声介导的Rac1基因沉默可能成为抑制结肠癌细胞侵袭和转移的一种新的治疗选择。
